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Quantification of Infectious Sendai Virus Using Plaque Assay

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Quantification of Infectious Sendai Virus Using Plaque Assay

Narihito Tatsumoto et al. Bio Protoc.

Abstract

Sendai virus (SeV) is an enveloped, single-stranded RNA virus of the family Paramyxoviridae. SeV is a useful tool to study its infectious pathomechanism in immunology and the pathomechanism of a murine model of IgA nephropathy. Virus quantification is essential not only to determine the original viral titers for an appropriate application, but also to measure the viral titers in samples from the harvests from experiments. There are mainly a couple of units/titers for Sendai viral quantification: plaque-forming units (PFU) and hemagglutination (HA) titer. Of these, we here describe a protocol for Sendai virus plaque assay to provide PFU using LLC-MK2 cells (a rhesus monkey kidney cell lines) and Guinea pig red blood cells. This traditional protocol enables us to determine Sendai virus PFU in viral stock as well as samples from your experiments.

Keywords: PFU; Paramyxivirus; Plaque assay; Plaque-forming units; Sendai virus; Titration.

Conflict of interest statement

Competing interests The authors do not have any potential conflicts of interest or competing interests to declare.

Figures

Figure 1.
Figure 1.. Lightly confluent LLC-MK2 cells.
Once overlaid agar is solidified, cells can no longer proliferate. Therefore, the cells want to be confluent when you overlay agar on the cells (Scale bar = 100 μm).
Figure 2.
Figure 2.. Serial dilutions of viral sample.
A. For whole-log dilutions, 100 μl of sample will be serially transferred to the next tube that contains 900 μl of pure Medium 199. B. For half-log dilutions, 316 μl of sample will be serially transferred to the next tube that contains 684 μl of pure Medium 199.
Figure 3.
Figure 3.. Sendai virus plaques.
A. Five plaques in one well. B. Magnification of one plaque shows a ring-shape plaque of red blood cells attaching to LLC-MK2 cells and a central empty area where LLC-MK2 cells died and detached.

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